Cardioprotective effect of tetra(aniline) containing terpolymers through miR-15a-5p and MFN-2 regulation against hypertrophic responses.

OBJECTIVE: Cardiac hypertrophy is a condition of abnormal cardiomyocyte enlargement accompanied by ventricular wall thickening. The study aims to investigate the role of miR-15a-5p in the regulation of mitofusin-2 (MFN-2) and to explore the cardioprotective effect of terpolymers ES-37 and L-37. METHODS: In this study, the Sprague Dawley rats' cardiac hypertrophic model was established by administering 5 mg/kg Isoproterenol subcutaneously every other day for 14 days. As treatment rats received NAC (50 mg/kg), NAC treatment (50 mg/kg NAC + 5 mg/kg ISO), ES-37 (1 mg/kg) and ES-37 treatment (1 mg/kg ES-37+5 mg/kg ISO), L-37 (1 mg/kg) and L-37 treatment (1 mg/kg L-37+5 mg/kg ISO). subcutaneously every other day for 14 days. NAC, ES 37 and L-37 were given after 1 h of Isoproterenol administration in treatment groups. Cardiac hypertrophy was confirmed through morphological and histological analysis. For estimation of oxidative stress profiling, ROS and TBARS and antioxidative profiling superoxide dismutase (SOD), Catalase, and Glutathione (GSH) levels were checked. Triglyceride, cholesterol, alanine transaminase (ALT), and aspartate transaminase (AST) were performed to evaluate levels of lipid profiling and liver profiling. Molecular expression analysis was checked through real-time PCR, and western blotting both at the transcriptional and translational levels. Molecular docking studies were performed to study the interactions and modes of binding between the synthetic polymers with three proteins (Mitofusin-2, DRP-1 and PUMA). All the studies were carried out using the AutoDock Vina software and the protein-ligand complexes were visualized in Biovia Discovery Studio. Cardiac hypertrophy was confirmed by the relative changes in the cellular structure of the heart by histopathological examination and physiological changes by estimating organ weights. Biochemical profiling results depict elevated oxidative and lipid profiles signify myocardial damage. N-acetyl cysteine (NAC), ES-37, and L-37 overcome the cardiac hypertrophic responses through attenuating oxidative stress and enhancing the antioxidative signaling mechanism. miR-15a-5p was identified as hypertrophic microRNA directly regulating the expression of Mitofusin-2 (MFN-2). Significantly increased expression of miR-15a-5p, Dynamin related protein 1 (Drp1), and P53 upregulated modulator of apoptosis (PUMA), was observed in the disease group, whereas MFN-2 expression was observed downregulated. N-acetyl cysteine (NAC), ES-37, and L-37 showed increased expression of antiapoptotic maker MFN-2 and decreased expression of miR-15a-5p, Drp1, and PUMA in treatment groups suggesting their cardioprotective role in attenuation of cardiac hypertrophy. An analysis of the docking results shows that ES-37 has greater binding affinity with the target proteins compared to L-37, with the highest binding values reported for MFN-2. CONCLUSION: The physiochemical properties of ES-37 and L-37 predicted it as a good drug-like molecule and its mechanism of action is predictably through inhibition of ROS. Molecular docking results shows that the polymer ES-37 has greater binding affinity with the target proteins compared to L-37, with the highest binding values reported for MFN-2. Thus, the study validates the role and targeting of miR-15a-5p and MFN-2 in cardiac hypertrophy as well as the therapeutic potential of NAC, ES-37, and L-37 in overcoming oxidative stress and myocardial damage.

N-Acetyl Cysteine, Selenium, and Ascorbic Acid Rescue Diabetic Cardiac Hypertrophy via Mitochondrial-Associated Redox Regulators.

Metabolic disorders often lead to cardiac complications. Metabolic deregulations during diabetic conditions are linked to mitochondrial dysfunctions, which are the key contributing factors in cardiac hypertrophy. However, the underlying mechanisms involved in diabetes-induced cardiac hypertrophy are poorly understood. In the current study, we initially established a diabetic rat model by alloxan-administration, which was validated by peripheral glucose measurement. Diabetic rats displayed myocardial stiffness and fibrosis, changes in heart weight/body weight, heart weight/tibia length ratios, and enhanced size of myocytes, which altogether demonstrated the establishment of diabetic cardiac hypertrophy (DCH). Furthermore, we examined the expression of genes associated with mitochondrial signaling impairment. Our data show that the expression of PGC-1α, cytochrome c, MFN-2, and Drp-1 was deregulated. Mitochondrial-signaling impairment was further validated by redox-system dysregulation, which showed a significant increase in ROS and thiobarbituric acid reactive substances, both in serum and heart tissue, whereas the superoxide dismutase, catalase, and glutathione levels were decreased. Additionally, the expression levels of pro-apoptotic gene PUMA and stress marker GATA-4 genes were elevated, whereas ARC, PPARα, and Bcl-2 expression levels were decreased in the heart tissues of diabetic rats. Importantly, these alloxan-induced impairments were rescued by N-acetyl cysteine, ascorbic acid, and selenium treatment. This was demonstrated by the amelioration of myocardial stiffness, fibrosis, mitochondrial gene expression, lipid profile, restoration of myocyte size, reduced oxidative stress, and the activation of enzymes associated with antioxidant activities. Altogether, these data indicate that the improvement of mitochondrial dysfunction by protective agents such as N-acetyl cysteine, selenium, and ascorbic acid could rescue diabetes-associated cardiac complications, including DCH.

Mentha longifolia Alleviates Exogenous Serotonin-Induced Diabetic Hypoglycemia and Relieves Renal Toxicity via ROS Regulation.

Diabetic kidney disease is one of the most common microvascular complications of diabetes mellitus with consequences of diabetic nephropathy. Here we amined to evaluate the nephroprotective potential of methanolic Mentha longifolia (MML) against serotonin-induced hypoglycemia allied toxicity in the rat model of diabetes. Diabetes was induced in rats via alloxan administration and validated by blood glucose level measurement. After that, the animals were treated with serotonin and methanolic extract of Mentha longifolia. Surprisingly, serotonin treatment significantly reduced the glucose levels to hypoglycemic conditions, accompanied by impaired redox defense system, abnormal kidney histopathology, dyslipidemia, and altered level of liver toxicity markers. Interestingly these changes were rescued by the methanolic extract of M. longifolia. The present study suggests that impaired serotonin levels during diabetic conditions may accelerate hypoglycemic allied free radical-dependent abnormalities; however, medicinal plants like M. longifolia can reduce these deleterious effects by scavenging free radicals and their associated toxicity.

Pistacia integerrima alleviated Bisphenol A induced toxicity through Ubc13/p53 signalling.

Exposure to environmental toxicants such as Bisphenol A (BPA) has raised serious health issues globally particularly in developing countries. It is ubiquitously used in the manufacturing of canned food and feeding bottles. BPA generated reactive oxygen species can lead to several diseases including cardiotoxicity. However, the endpoints stimulated in BPA cardiotoxicity yet need to be investigated. The current study was aimed to investigate the underlying molecular pathways which may contribute in revealing the protective effects of Pistacia integerrima against BPA induced oxidative stress. The dose of 100 µg/kg BW of BPA, 200 mg/kg BW P. integerrima, and 4 mg/kg BW melatonin was administered to Sprague Dawley rats. Present results of western blotting and qRT-PCR showed the increased expression of p53, PUMA and Drp1, while downregulation of Ubc13 in heart tissues of BPA treated group whereas the levels were reversed upon treatment with P. integerrima. The role of BPA in heart tissue apoptosis was further confirmed by the increased level of P-p53, cytochrome C and disrupted cellular architecture whereas the P. integerrima has shown its ameliorative potential by mitigating the adverse effects of BPA. Moreover, the oxidant, antioxidant, lipid, and liver markers profile has also revealed the therapeutic potential of P. integerrima by maintaining the levels in the normal range. However, melatonin has also manifested the normalized expression of apoptotic markers, biochemical markers, and tissue architecture. Conclusively, the data suggest that P. integerrima may be a potential candidate for the treatment of BPA induced toxicity by neutralizing the oxidative stress through Ubc13/p53 pathway.

An Overview of Non-coding RNAs and Cardiovascular System.

Cardiovascular disease management and timely diagnosis remain a major dilemma. Delineating molecular mechanisms of cardiovascular diseases is opening horizon in the field of molecular medicines and in the development of early diagnostic markers. Non-coding RNAs are the highly functional and vibrant nucleic acids and are known to be involved in the regulation of endothelial cells, vascular and smooth muscles cells, cardiac metabolism, ischemia, inflammation and many processes in cardiovascular system. This chapter is comprehensively focusing on the overview of the non-coding RNAs including their discovery, generation, classification and functional regulation. In addition, overview regarding different non-coding RNAs as long non-coding, siRNAs and miRNAs involvement in the cardiovascular diseases is also addressed. Detailed functional analysis of this vast group of highly regulatory molecules will be promising for shaping future drug discoveries.

Prospective Advances in Non-coding RNAs Investigation.

Non-coding RNAs (ncRNAs) play significant roles in numerous physiological cellular processes and molecular alterations during pathological conditions including heart diseases, cancer, immunological disorders and neurological diseases. This chapter is focusing on the basis of ncRNA relation with their functions and prospective advances in non-coding RNAs particularly miRNAs investigation in the cardiovascular disease management.The field of ncRNAs therapeutics is a very fascinating and challenging too. Scientists have opportunity to develop more advanced therapeutics as well as diagnostic approaches for cardiovascular conditions. Advanced studies are critically needed to deepen the understanding of the molecular biology, mechanism and modulation of ncRNAs and chemical formulations for managing CVDs.

Interplay of N acetyl cysteine and melatonin in regulating oxidative stress-induced cardiac hypertrophic factors and microRNAs.

Early and specific diagnosis of oxidative stress linked diseases as cardiac heart diseases remains a major dilemma for researchers and clinicians. MicroRNAs may serve as a better tool for specific early diagnostics and propose their utilization in future molecular medicines. We aimed to measure the microRNAs expressions in oxidative stress linked cardiac hypertrophic condition induced through stimulants as Endothelin and Isoproterenol. Cardiac hypertrophic animal models were confirmed by BNP, GATA4 expression, histological assays, and increased cell surface area. High oxidative stress (ROS level) and decreased antioxidant activities were assessed in hypertrophied groups. Enhanced expression of miR-152, miR-212/132 while decreased miR-142-3p expression was observed in hypertrophic condition. Similar pattern of these microRNAs was detected in HL-1 cells treated with H2O2. Upon administration of antioxidants, the miRNAs expression pattern altered from that of the cardiac hypertrophied model. Present investigation suggests that oxidative stress generated during the cardiac pathology may directly or indirectly regulate anti-hypertrophy pathway elements through microRNAs including antioxidant enzymes, which need further investigation. The down-regulation of free radical scavengers make it easier for the oxidative stress to play a key role in disease progression.

Customer Prioritization Integrated Supply Chain Optimization Model with Outsourcing Strategies.

Pre-COVID-19, most of the supply chains functioned with more capacity than demand. However, COVID-19 changed traditional supply chains' dynamics, resulting in more demand than their production capacity. This article presents a multiobjective and multiperiod supply chain network design along with customer prioritization, keeping in view price discounts and outsourcing strategies to deal with the situation when demand exceeds the production capacity. Initially, a multiperiod, multiobjective supply chain network is designed that incorporates prices discounts, customer prioritization, and outsourcing strategies. The main objectives are profit and prioritization maximization and time minimization. The introduction of the prioritization objective function having customer ranking as a parameter and considering less capacity than demand and outsourcing differentiates this model from the literature. A four-valued neutrosophic multiobjective optimization method is introduced to solve the model developed. To validate the model, a case study of the supply chain of a surgical mask is presented as the real-life application of research. The research findings are useful for the managers to make price discounts and preferred customer prioritization decisions under uncertainty and imbalance between supply and demand. In future, the logic in the proposed model can be used to create web application for optimal decision-making in supply chains.

Melatonin abated Bisphenol A-induced neurotoxicity via p53/PUMA/Drp-1 signaling.

Bisphenol A (BPA), an endocrine disruptor, is widely used in the manufacture of different daily life products. Accumulating evidence supports the association between the increasing incidence of neurodegenerative diseases and the BPA level in the environment. In the present study, we aimed to evaluate the neuroprotective role of melatonin against BPA-induced mitochondrial dysfunction-mediated apoptosis in the brain. Herein, adult Sprague Dawley rats were administrated (subcutaneously) with BPA (100 µg/kg BW, 1 mg/kg BW, and 10 mg/kg BW) and melatonin (4 mg/kg BW) for 16 days. Our results showed BPA exposure significantly increased the oxidative stress as demonstrated by increased free radicals (ROS), TBARs level, disrupted cellular architecture, and decreased antioxidant enzymes including SOD, CAT, APX, POD, and GSH levels. Additionally, BPA treatment increased the expression of PUMA, p53, and Drp-1 resulting in apoptosis in the brain tissue of rats. However, melatonin treatment significantly attenuated BPA-induced toxic effects by scavenging ROS, boosting antioxidant enzyme activities, and interestingly enervated brain apoptosis by normalizing p53, PUMA, and Drp-1 expressions at both transcriptional and translational level. Moreover, the brain tissue histology also revealed the therapeutic potential of melatonin by normalizing the cellular architecture. Conclusively, our finding suggests that melatonin could alleviate oxidative stress and mitochondrial dysfunction-linked apoptosis, rendering its neuroprotective potential against BPA-induced toxicity.

Engineering electroactive and biocompatible tetra(aniline)-based terpolymers with tunable intrinsic antioxidant properties in vivo.

Under different pathological conditions, high levels of reactive oxygen species (ROS) cause substantial damage to multiple organs. To counter these ROS levels in multiple organs, we have engineered highly potent novel terpolymers. We found that combination of FDA-approved polyethylene glycol, fumaric acid moieties and electroactive tetra(aniline) by varying the content of tetra(aniline) results into a novel drug composition with biologically active and tunable intrinsic antioxidant properties. To test the intrinsic antioxidative properties of these novel terpolymers, we used alloxan to induce diabetes in rats where ROS generation is known to be higher. The systemic administration of terpolymers to the diabetic rats showed strong electroactive antioxidant behavior which not only normalized ROS levels, but also improved the levels of enzymatic antioxidants including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). As a proof-of-principle, we here show TANI based novel drug composition of terpolymers with tunable intrinsic antioxidant properties in multiple organs.

Giant cell granuloma of the maxilla.

The case report is of a 30 year old male, who complained of a slowly expanding swelling in the maxilla in the left premolar region. The swelling was clinically and radiographically diagnosed as Central Giant Cell Granuloma. The patient also had Angles Class 2 Division 2 Malocclusion in which repeated micro trauma may have possibly contributed to the formation of the tumor. The tumor was surgically removed under general anesthesia followed by local injections of corticosteroids.

Pathomorphological alterations associated with chicken coccidiosis in Jammu division of India.

Pathomorphological alterations of chicken coccidiosis in Jammu division were undertaken in both organized and backyard chickens during the year 2010-2011. A total 240 intestines were examined from both organized farms and backyard chickens for histopathological studies. Out of 240 samples processed, 48 samples were found to be positive for coccidiosis with a prevalence of 20 %. Coccidiosis was initially diagnosed on the basis of faecal examination and confirmed by the presence of sporulated oocysts and pathomorphological alterations in intestines. Eimeria species were identified by morphometry. Five Eimeria species identified were Eimeria tenella, E. necatrix, E. maxima, E. acervulina and E. mitis. Histopathological lesions revealed loss of epithelial tissue, congestion of blood vessels which indicate disruption followed by haemorrhage, severe muscular oedema and necrosis of submucosa of intestine and caecum. There was loss of intestinal villi, disruption of caecal mucosa and clusters of oocysts seen. There was massive infiltration by heterophils and mononuclear cells. Several merozoites, schizonts and microgametes were found in the epithelial cells.

Treatment of mange infection in a weaner flock of sheep with ivermectin at sheep breeding farm Hardishiva of Kashmir valley.

An outbreak of Sarcoptes scabiei mange occured in a weaner flock that comprised of 74 animals including 31 male and 43 female at sheep breeding farm Hardishiva of Kashmir valley. All the animals in the flock were affected. Animals were very weak and emaciated skin lesions were confined to the lips, nostrils, ears, face, and were characterized by pruritus, erythema, papule formation, alopecia and scab formation. Microscopic examination of skin scraping revealed S. scabiei mange infestation. The affected animals were treated with ivermectin at 1 ml/50 kg b.wt. S/C, as single dose along with supportive therapy. 7th day post treatment scrapping examinations revealed absence of mites or their eggs in these scrapings.

Prevalence of poultry coccidiosis in Jammu region of Jammu & Kashmir State.

In this study prevalence of chicken coccidiosis in Jammu division were undertaken in both organized and backyard chickens during the year 2010-2011, with an overall prevalence of 39.58 % on examination of 720 faecal samples. Five Eimeria species were identified viz., E. tenella, E. necatrix, E. maxima, E. acervulina and E. mitis. E. tenella was the predominant species in both organized and unorganized farms. The highest prevalence percentage was found in July, 2011 (68.9 %) and the lowest percentage was found in May, 2011 (12.5 %). Coccidial prevalence was found to be 53.61 % in unorganized (backyard poultry birds) as compared to organized birds (25.55 %). Maximum positive cases of coccidian infection was found in monsoon season (60.55 %) and least in summer season (21.66 %). Birds of age 31-45 days showed more prevalence percentage (58.86 %). Higher oocysts count was recorded from July to September with a peak value (38973.00 ± 3075.6) in July and lowest (12914.00 ± 595.48) in the month of May.

Evaluation of the Ochsenbein-Luebke flap technique in periapical surgery at Punjab Dental Hospital Lahore Pakistan.

BACKGROUND: There have been no direct studies comparing the Trapezoidal flap with the Ochsenbein-Luebke flap in endodontic surgery. The present study is aimed to provide an insight into the relative performances of these two procedures. METHODS: This comparative study, carried out at Punjab Dental Hospital Lahore Pakistan from May to October 1998, involved 120 patients (60 in each group). Evaluation of technique(s) involved assessment of intra-operative technical considerations as well as postoperative morbidity, healing and cosmetic results. RESULTS: It was found that O-L flap is easy to reflect, has good visibility, less bleeding duration and is easy to handle during surgery. Stitches are less time consuming, there is less tearing, better cosmetic result and no recession of gingiva. However more studies should be done to evaluate the procedure in larger number of patients.